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Structural, functional and chemical changes in Pseudozyma antarctica lipase B on exposure to hydrogen peroxide.

Törnvall, Ulrika LU ; Hedström, Martin LU ; Schillén, Karin LU and Hatti-Kaul, Rajni LU (2010) In Biochimie 92(Online 21 July 2010). p.1867-1875
Abstract
The effect on primary, secondary, tertiary and quaternary structure of Pseudozyma (formerly Candida) antarctica lipase B (PalB) on exposure to hydrogen peroxide was investigated using nano-electrospray ionization-mass spectrometry (nano-ESI-MS), liquid chromatography tandem mass spectrometry (LC/MS/MS), circular dichroism (CD), and dynamic light scattering (DLS). Treatment with hydrogen peroxide generated heavier protein variants, with a mass gain that increased with increasing incubation time. Furthermore, elevated concentration of H(2)O(2) was shown to result in partial fragmentation of the protein. Proteolytic digestion of the enzyme gave primary sequence coverage of more than 90%, revealing oxidation of methionine, tryptophan and... (More)
The effect on primary, secondary, tertiary and quaternary structure of Pseudozyma (formerly Candida) antarctica lipase B (PalB) on exposure to hydrogen peroxide was investigated using nano-electrospray ionization-mass spectrometry (nano-ESI-MS), liquid chromatography tandem mass spectrometry (LC/MS/MS), circular dichroism (CD), and dynamic light scattering (DLS). Treatment with hydrogen peroxide generated heavier protein variants, with a mass gain that increased with increasing incubation time. Furthermore, elevated concentration of H(2)O(2) was shown to result in partial fragmentation of the protein. Proteolytic digestion of the enzyme gave primary sequence coverage of more than 90%, revealing oxidation of methionine, tryptophan and cystine residues. The active site histidine was not observed in oxidized form in any of the experiments. However, oxidation of cystine to cysteic acid indicated disruption of disulphide bridges, and CD evaluations confirmed that severe changes to the secondary structure towards random coil had occurred. The structural changes could be an effect of the observed amino acid side chain oxidations, and was correlated with deactivation of the lipase. From DLS experiments, it was seen that the lipase exposed to both high temperature and H(2)O(2) formed large and intermediate sized aggregates, not observed for the heat treated enzyme. The findings reported here could lay the basis for developing enzyme variants with higher oxidative stability. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochimie
volume
92
issue
Online 21 July 2010
pages
1867 - 1875
publisher
Elsevier
external identifiers
  • wos:000286295900021
  • pmid:20654682
  • scopus:78650232067
ISSN
1638-6183
DOI
10.1016/j.biochi.2010.07.008
language
English
LU publication?
yes
id
947ed59d-ce4e-40ae-a8a0-5510935c36d6 (old id 1644599)
date added to LUP
2010-08-23 12:56:25
date last changed
2018-05-29 11:08:58
@article{947ed59d-ce4e-40ae-a8a0-5510935c36d6,
  abstract     = {The effect on primary, secondary, tertiary and quaternary structure of Pseudozyma (formerly Candida) antarctica lipase B (PalB) on exposure to hydrogen peroxide was investigated using nano-electrospray ionization-mass spectrometry (nano-ESI-MS), liquid chromatography tandem mass spectrometry (LC/MS/MS), circular dichroism (CD), and dynamic light scattering (DLS). Treatment with hydrogen peroxide generated heavier protein variants, with a mass gain that increased with increasing incubation time. Furthermore, elevated concentration of H(2)O(2) was shown to result in partial fragmentation of the protein. Proteolytic digestion of the enzyme gave primary sequence coverage of more than 90%, revealing oxidation of methionine, tryptophan and cystine residues. The active site histidine was not observed in oxidized form in any of the experiments. However, oxidation of cystine to cysteic acid indicated disruption of disulphide bridges, and CD evaluations confirmed that severe changes to the secondary structure towards random coil had occurred. The structural changes could be an effect of the observed amino acid side chain oxidations, and was correlated with deactivation of the lipase. From DLS experiments, it was seen that the lipase exposed to both high temperature and H(2)O(2) formed large and intermediate sized aggregates, not observed for the heat treated enzyme. The findings reported here could lay the basis for developing enzyme variants with higher oxidative stability.},
  author       = {Törnvall, Ulrika and Hedström, Martin and Schillén, Karin and Hatti-Kaul, Rajni},
  issn         = {1638-6183},
  language     = {eng},
  number       = {Online 21 July 2010},
  pages        = {1867--1875},
  publisher    = {Elsevier},
  series       = {Biochimie},
  title        = {Structural, functional and chemical changes in Pseudozyma antarctica lipase B on exposure to hydrogen peroxide.},
  url          = {http://dx.doi.org/10.1016/j.biochi.2010.07.008},
  volume       = {92},
  year         = {2010},
}